The present invention relates to a carrier for affinity chromatgraphy.
The rapid development of biotechnology enables production of various materials having value different from conventional industrial techniques. Namely, with the development of genetic engineering, cell fusion techniques and the like, many techniques for production of useful materials by using their biological function are known. The efficient technique of high purification is a most important matter in laboratories and in factories. For example, gene manipulation has enabled production of trace amounts of physiological active materials such as hormones, lymphokines and the like on an industrial level. However, separation and purification methods involving efficiently and stably separating these materials and removing impurities are desired. As such separation and purification methods, gel filtration, ion exchange and the like have been utilized. These methods need operations having many steps for obtaining materials of adequate purity in high cost and in low yields.
Fundamentally, it is considered to fit the purpose that specific molecular recognition in a cell is used for the separation of such materials which develop physiological function in vivo.
By using specific affinity between special biochemical materials such as an enzyme and an inhibitor an, enzyme and a substrate, an antigen and an antibody a, hormone and a receptor and the like, affinity chromatography can separate these materials. Thus, the technique of affinity chromatography becomes widely used to separate and purify these materials. As the technique using the reaction between an antigen and an antibody, there is immuno affinity chromatography characterized in that the biological affinity specificity is very high. According to this technique, the desired materials to be purified are selectively adsorbed on adsorbent which is obtained by binding an antibody for the desired material as a ligand to an insoluble carrier. The immuno affinity chromatography is applicable to protein, glycoprotein, peptide, enzyme, peptide type hormone and the like, antibodies of which can be produced.
In conventional immuno affinity chromatography, antibodies of glycoprotein are drawn from blood in which antigens are immunized or introduced in mammals such as rabbits, horses, cows, and goats, the resulting antibodies are purified by gel filtration, protein A affinity chromatography and the like, and the antibodies are immobilized on an insoluble substrate. Then, through the specific binding ability between protein as the antibodies and the antigens, the antigens are purified by an immunological method. As the binding of the antibodies and the antigens is comparatively strong, the complex obtained by the reaction is stable. The binding is reversible, and the antigens are dissociated from the immobilized antibodies and purified by changing their conditions.
The antibodies which are used in the above described conventional techniques are protein. The sample liquids containing the antigens mostly contain proteolytic enzyme (protease). For this reason, immobilized antibodies are degraded by the protease, and the binding activity of the antigen of the afinity chromatography is lost. Antibodies are frequently available only at high prices or in slight amounts. It becomes important to use these antibodies efficiently, i.e., continuously without their inactivation.